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丙型肝炎4a基因型核心蛋白基因的鉴定、克隆与表达

Identification, cloning and expression of core protein gene of hepatitis C genotype 4a.

作者信息

Sabet Salwa F, Al-Sherbiny Maged M, Hagan Paul

机构信息

Department of Zoology, Faculty of Science, Cairo University.

出版信息

J Egypt Soc Parasitol. 2009 Dec;39(3):769-87.

Abstract

Although Egypt has very high rates of HCV, not much is known about genotype 4a which is the most predominant genotype in Egypt. In the present study, core (C_ED43) gene of the Egyptian strain ED43 of HCV genotype 4a was first analyzed using PC/GENE program. Computer analysis of Core region of the isolate ED43 revealed that the Egyptian genotype 4a is different from those isolated from Europe and Central Africa and that it is closely related to genotype 1b. The DNA region coding for the Core was amplified from HCV_ED43/PUC19 plasmid. The PCR product was then cloned and expressed in E. coli M15 using pQE-30 vector. The expression and antigenicity of the core (Core_4a) protein in E. coli was confirmed by SDS-PAGE and western blotting, which will make it useful for developing assay systems for detecting anti-HCV antibodies and HCV antigen, respectively and which might help in the design of a vaccine against the Egyptian genotype 4a.

摘要

尽管埃及丙型肝炎病毒(HCV)感染率很高,但对于埃及最主要的基因型4a,人们了解并不多。在本研究中,首先使用PC/GENE程序分析了HCV基因型4a埃及毒株ED43的核心(C_ED43)基因。对分离株ED43核心区域的计算机分析表明,埃及基因型4a与从欧洲和中非分离出的毒株不同,且与基因型1b密切相关。从HCV_ED43/PUC19质粒中扩增出编码核心的DNA区域。然后将PCR产物克隆,并使用pQE - 30载体在大肠杆菌M15中表达。通过SDS - PAGE和蛋白质印迹法证实了核心(Core_4a)蛋白在大肠杆菌中的表达和抗原性,这将使其分别有助于开发检测抗HCV抗体和HCV抗原的检测系统,并且可能有助于设计针对埃及基因型4a的疫苗。

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